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KSR1 and cPLA2 localize at the PM of wild-type (wt) but not H/N-Ras−/− MEFs upon stimulation. Soluble (S100) and particulate (P100) fractions were isolated from wild-type and H/N-Ras−/− MEFs, starved (−) or stimulated (+) with EGF (10 ng/ml, 5 min). The presence of KSR1 and cPLA2 was examined by immunoblotting. The purity of the fractions was ascertained by blotting against Rho-GDP dissociation inhibitor (αGDI) and transferrin (Tfr) receptor in lysates from wild-type MEFs.
